Murine Model of Metastatic Liver Tumors in the Setting of Ischemia Reperfusion Injury

Summary

We describe in detail a clinically relevant colorectal cancer liver metastases (CRLM) tumor model and the influence of liver ischemia reperfusion (I/R) in tumor growth and metastasis. This model can help to better understand the mechanisms underlying surgery-induced promotion of liver metastatic growth.

Abstract

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce multiple tissue and organ damage. Despite recent advances and therapeutic approaches, the overall morbidity has remained unsatisfactory especially in patients with underlying parenchymal abnormalities. In the context of aggressive cancer growth and metastasis, surgical I/R is suspected to be the promoter regulating tumor recurrence. This article aims to describe a clinically relevant murine model of liver I/R and colorectal liver metastasis. In doing so, we aim to assist other investigators in establishing and perfecting this model for their routine research practice to better understand the effects of liver I/R on promoting liver metastases.

Introduction

The liver is one of the most common sites for the development of metastatic disease¹. Mortality is almost invariably attributable to complications associated with tumor growth in the liver. In patients with metastatic solid tumors in the liver, surgery remains a crucial intervention for disease control and a possible curative approach. However, the vast majority of patients ultimately present with recurrent disease, predominantly in the liver^2,3. During hepatic surgery, intraoperative bleeding is common, often necessitating blood transfusion and different technical approaches for contro....

Protocol

All animal protocols are approved by the Institutional Animal Care and Use Committee and adhered to the National Institutes of Health (NIH) Guidelines. Instruments used for any surgical procedure were thoroughly sterilized.

1. Initial preparation

1. Before injecting cancer cells into the mouse spleen, autoclave and sterilize all instruments to be used during the procedure.
2. Sterilize and/or autoclave a heating pad, surgical gloves, gauze, pairs of scissors, small clamps, vess.......

Discussion

The animal model described in this manuscript is based upon two major approaches. The first is to recognize the ability of cancer cells to localize and proliferate in the liver lobes. The second is to study the effect of hepatic ischemia reperfusion injury influencing the tumor growth and metastases. This model permits the relevant study of liver metastases in the absence of secondary metastases in an immunocompetent mouse. The model is useful in addressing the questions of metastatic efficiency, such as cell survival ex.......

Materials

Name	Company	Catalog Number	Comments
Dulbecco's Modified Eagle Medium	Lonza	12-614F
Fetal Bovine Serum	Lonza	900-108
L-Glutamin	Gibco	25030-081
Penicilin	Fisher scientific	15-140-122
Stretomysin	Fisher scientific	15-140-122
HEPES	Fisher Scientific	SH3023701
Trypsin	Hyclone	sh30042.02
Cell culture Flask 75cm	5 Cells Star	658170
15ml PP Conical Tubes	BioExcell	41021037
Trypan Blue Stain	Giibco	15250-061
Gauze	Fisherbrand	1376152
Cautry	Bovie	AA01
Microvascular clamp	Finescience tools	18055-03
Micro-Serrefine clamp applicator with lock	Fine science toosl	FST-18056-14
Spring scissor	Fine science toosl	FST-15021-15
Vessel Dilator	Fine science toosl	FST-00276-13
Magnetic fixator Retraction system	Fine science toosl	FST-18200020
Micro-Adson Forceps	Fine science toosl	FST-11019-12
Micro-Adson Forceps	Fine science toosl	FST-11018-12
4-0 polypropylene suture	Ethicon	K881H
Needle holder	Harvard Apparatus	72-8826
Heating Pad	Fisher scientific	1443915
Clipper	Oster	559A
Povidone-Iodine solution	Medline	MDS093945
Syringe 1ml 25G	BD safety Glide	305903
Insulin syringe 0.5 ml	BD insulin Syringes	32946
Cotton -Tipped Applicator	Fisher Scientific	23-400-101
Surgical Microscope	Leica	LR92240
Mycoplasma Elisa Kit	Roche	11663925910
Ketamine	Putney	#056344
Xylazine	NADA	#139-236
ALT strip	Heska	15809554
AST strip	Heska	15809542
LDH strip	Heska	15809607